Geerts Pascal, Toussaint André, Mergeai Guy, Baudoin Jean-Pierre
Life Science Department, Walloon Agricultural Research Center, Gembloux, Belgium.
Methods Mol Biol. 2011;710:117-29. doi: 10.1007/978-1-61737-988-8_10.
Predominant among the production constraints of the common bean Phaseolus vulgaris are infestation of Ascochyta blight, Bean Golden Mosaic virus (BGMV), and Bean Fly. Interbreeding with Phaseolus -coccineus L. and/or Phaseolus polyanthus Greenm has been shown to provide P. vulgaris with greater resistance to these diseases. For interspecific crosses to be successful, it is important to use P. coccineus and P. polyanthus as female parents; this prevents rapid reversal to the recurrent parent P. vulgaris. Although incompatibility barriers are post-zygotic, early hybrid embryo abortion limits the success of F1 crosses. While rescue techniques for globular and early heart-shaped embryos have improved in recent years, -success in hybridization remains very low. In this study, we describe six steps that allowed us to rescue 2-day-old P. vulgaris embryos using a pod culture technique. Our methods consisted of (i) pod culture, (ii) extraction and culture of immature embryos, (iii) dehydration of embryos, (iv) germination of embryos, (v) rooting of developed shoots, and (vi) hardening of plantlets.
普通菜豆(Phaseolus vulgaris)生产受限的主要因素是受到壳二孢叶枯病、菜豆金色花叶病毒(BGMV)和豆蝇的侵害。与多花菜豆(Phaseolus coccineus L.)和/或柔毛菜豆(Phaseolus polyanthus Greenm)杂交已被证明能使普通菜豆对这些病害具有更强的抗性。为使种间杂交成功,将多花菜豆和柔毛菜豆用作母本很重要;这可防止迅速回复到轮回亲本普通菜豆。虽然不亲和障碍是合子后障碍,但早期杂交胚败育限制了F1杂交的成功率。尽管近年来针对球形胚和早期心形胚的拯救技术有所改进,但杂交成功率仍然很低。在本研究中,我们描述了六个步骤,通过荚培养技术拯救2日龄的普通菜豆胚。我们的方法包括:(i)荚培养,(ii)未成熟胚的提取和培养,(iii)胚脱水,(iv)胚萌发,(v)发育芽生根,以及(vi)试管苗硬化。